Hoseini Seyed Mehdi, Montazeri Fateme, Moghaddam-Matin Maryam, Bahrami Ahmad Reza, Meimandi Hassan Heidarian, Ghasemi-Esmailabad Saeed, Kalantar Seyed Mehdi
Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran.
Biotechnology Research Center, International Campus, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran.
Int J Reprod Biomed. 2020 Oct 13;18(10):885-898. doi: 10.18502/ijrm.v13i10.7773. eCollection 2020 Oct.
The genomic stability of stem cells to be used in cell therapy and other clinical applications is absolutely critical. In this regard, the relationship between in vitro expansion and the chromosomal instability (CIN), especially in human amniotic fluid cells (hAFCs) has not yet been completely elucidated.
To investigate the CIN of hAFCs in primary and long-term cultures and two different culture mediums.
After completing prenatal genetic diagnoses (PND) using karyotype technique and chromosomal analysis, a total of 15 samples of hAFCs from 650 samples were randomly selected and cultured in two different mediums as AmnioMAX II and DMEM. Then, proliferative cells were fixed on the slide to be used in standard chromosome G-banding analysis. Also, the senescent cells were screened for aneuploidy considering 8 chromosomes by FISH technique using two probe sets including PID I (X-13-18-21) & PID II (Y-15-16-22).
Karyotype and interphase fluorescence in situ hybridization (iFISH) results from 650 patients who were referred for prenatal genetic diagnosis showed that only 6 out of them had culture- derived CIN as polyploidy, including mosaic diploid-triploid and diploid-tetraploid. Moreover, the investigation of aneuploidies in senesced hAFCs demonstrated the rate of total chromosomal abnormalities as 4.3% and 9.9% in AmnioMAX- and DMEM-cultured hAFCs, respectively.
hAFCs showed a low rate of CIN in two AmnioMAX II and DMEM mediums and also in the proliferative and senescent phases. Therefore, they could be considered as an attractive stem cell source with therapeutic potential in regenerative medicine.
用于细胞治疗和其他临床应用的干细胞的基因组稳定性至关重要。在这方面，体外扩增与染色体不稳定性（CIN）之间的关系，尤其是在人羊水细胞（hAFCs）中的关系尚未完全阐明。
研究原代和长期培养以及两种不同培养基中hAFCs的CIN。
使用核型技术和染色体分析完成产前基因诊断（PND）后，从650个样本中随机选取15个hAFCs样本，在两种不同的培养基（AmnioMAX II和DMEM）中培养。然后，将增殖细胞固定在载玻片上用于标准染色体G显带分析。此外，使用包括PID I（X-13-18-21）和PID II（Y-15-16-22）的两个探针组，通过荧光原位杂交（FISH）技术筛选衰老细胞的非整倍体。
650例接受产前基因诊断患者的核型和间期荧光原位杂交（iFISH）结果显示，其中只有6例因培养产生CIN，表现为多倍体，包括嵌合二倍体 - 三倍体和二倍体 - 四倍体。此外，对衰老hAFCs非整倍体的研究表明，在AmnioMAX和DMEM培养的hAFCs中，总染色体异常率分别为4.3%和9.9%。
hAFCs在两种AmnioMAX II和DMEM培养基中以及增殖期和衰老期均显示出较低的CIN发生率。因此，它们可被视为再生医学中具有治疗潜力的有吸引力的干细胞来源。
